Method for Discovering Novel DNA Viruses in Blood Using Viral Particle Selection and Shotgun Sequencing | Zendy

Mya Breitbart | Zendy; Forest Rohwer | Zendy

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Method for Discovering Novel DNA Viruses in Blood Using Viral Particle Selection and Shotgun Sequencing

Author(s) -

Mya Breitbart,

Forest Rohwer

Publication year - 2005

Publication title -

biotechniques

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.617

H-Index - 131

eISSN - 1940-9818

pISSN - 0736-6205

DOI - 10.2144/000112019

Subject(s) - shotgun sequencing , shotgun , biology , computational biology , dna , selection (genetic algorithm) , virology , population , cloning (programming) , dna sequencing , polymerase chain reaction , human virome , genetics , genome , gene , computer science , medicine , artificial intelligence , programming language , environmental health

Rapid identification of viruses is needed to monitor the blood supply for emerging threats. Here we present a method that meets these criteria and allows for the shotgun sequencing of novel, uncultured DNA viruses directly from human blood. This method employs selection based on the physical properties of viruses combined with sequence-independent amplification and cloning. We show that both single- and double-stranded DNA viruses can be recovered from blood samples using this approach. In addition, we report the discovery of novel anellovirus sequences in the blood of healthy donors. PCR primers designed to amplify these novel anellovirus sequences were then used to verify the presence of these viruses in the general donor population.

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